1. Field of the Invention
The present invention generally involves polymeric materials and methods of using the polymeric materials for immobilizing biopolymers. More particularly, the present invention relates to activated polymeric films and molded parts, methods for preparing activated polymeric films and molded parts and methods for immobilizing biopolymers onto activated polymeric films and molded parts.
2. Description of Related Art
For many years solid phase chemistry has found a rapidly expanding utility in biopolymer synthesis and biopolymer immobilization. For example, organic and inorganic materials have been utilized for the solid phase synthesis of peptides and oligonucleotides by the step wise addition of activated amino acid derivatives or nucleotide derivatives to a growing oligomeric chain attached at one end to a solid support. Typically, the solid support material is in the form of a porous bead or resin material having a high surface area. Typically, in these solid phase systems, the chemically synthesized peptides or oligonucleotides are cleaved from the solid support and subsequently utilized in biochemical research or diagnostic applications. Thus, the solid support material plays no role in the research or diagnostic applications, having been used only during the initial synthesis of the biopolymer.
Additionally, solid phase chemistry has gained recognition for its usefulness in the study and the analysis of biopolymers. For example, cellulose membranes, polypropylene membranes, and nylon membranes have been widely used in blotting techniques which involve capturing native biopolymers onto the membrane and subsequently immobilizing the blotted biopolymer using heat, radiation or chemical techniques. The immobilized biopolymers are then available for subsequent analyses such as those associated with Southern blotting applications and reverse hybridization analytical techniques.
Additionally, presynthesized or natural oligonucleotides have been immobilized by covalently attaching activated oligonucleotides to the solid support. Typically, this approach requires activating the oligonucleotide with, e.g., a carbodiimide. Unfortunately, the activated oligonucleotides are expensive and they have short useful lives because they are very unstable. Thus, preparing and utilizing these activated oligonucleotides often lead to the loss of expensive reagents when the activated oligonucleotide decays to an inactive form. Moreover, carbodiimide activation frequently results in urea side product formation. Since these ureas tend to be insoluble in many common organic solvents their presence in automated reaction systems can cause problems when tubing and other lines are clogged by the precipitate.
More recently, oligonucleotides and peptides have been synthesized chemically to a solid support material directly and subsequently utilized without cleaving the biopolymers from the solid support. Generally, these support bound biopolymers are located at site specific locations on the solid support, with each site associated with at least one known biopolymer. Contacting, for example, site specifically bound oligonucleotides to unknown oligonucleotides under hybridizing conditions and subsequently determining the sites at which hybridization occurs can provide information relating to the structure and biological function of the unknown oligomer. Solid support materials suitable for such applications must physically and chemically withstand the temperatures and solvents associated with synthesizing the biopolymer onto the support, and not interfere with the hybridization and detection process.
Derivatized polypropylene films, glass slides and silicon wafers have been used for the solid support synthesis of oligonucleotides and peptides at site specific locations on the film, slide or wafer. These materials have been fairly successful because the glass, polypropylene and silicon withstand the physical and chemical rigors of the synthesis and hybridization processes. Furthermore, these materials are suitable when fluorescence detection techniques are used because they have low background fluorescence. However, glass slides, silicon wafers and polymer films are difficult to handle, and require handles or specially designed holders in order to manipulate the solid support when they are utilized in automated processes. These handles or holders are expensive to design and use and generally add to the costs of the utilizing solid supports. Furthermore, it is difficult to characterize and control the surface density of biopolymers synthesized or attached to glass slides and polypropylene films. Thus, there is a continuing need for improved materials suitable for immobilizing biopolymers and for materials suitable for directly synthesizing biopolymers. Such materials are preferably also suitable as solid supports for the evaluation of immobilized biopolymer biological activity, their identification or their use in analytical applications.
Accordingly, it is an object of the present invention to provide biopolymer immobilization reagents which are stable under conditions for synthesizing or immobilizing biopolymers and which are stable under conditions for the subsequent analysis of biopolymers.
It is also an object of the present invention to provide biopolymer immobilization reagents which are capable of being formed into devices which intrinsically incorporate holders or handlers.
It is a further object of the present invention to provide biopolymer immobilization reagents which are inexpensive and physically rugged.
It is another object of the present invention to provide biopolymer immobilization reagents whose compositions are controllable to provide greater or lesser surface densities of biopolymer synthesis.
It is another object of the present invention to provide reagents having surfaces suitable for immobilizing biopolymers at site specific locations and at high biopolymer densities.
It is also an object of the present invention to provide biopolymer immobilization reagents which utilize solid materials capable of forming films or flat planar surfaces and do not interfere with detection techniques.
It is further an object of the present invention to provide biopolymer immoblization reagents which have reasonable shelf lives and do not degrade at ambient conditions.